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为研究MHCⅡ类基因转录活化因子(CⅡTA)对人类白细胞抗原(HLA)Ⅱ类分子表达的影响,用RT-PCR方法从Raji细胞中克隆到C Ⅱ TA基因cDNA 5'端片段,构建成CⅡTA反义RNA真核表达载体 pcDNA-Ⅱ,基因转移 HLA Ⅱ类分子诱导型表达的 HeLa细胞.经 IFN-γ诱导,发现稳定转染pcDNA-Ⅱ的细胞中HLA-DR,DP和DQ的表达均较转pcDNA3空载体的细胞有显著下降,而 HLAⅠ类分子的表达不受影响.实验证明 CⅡ TA反义 RNA对 HLA Ⅱ类分子表达具有显著的抑制作用,为进一步开展低免疫原性的细胞移植提供了依据.
In order to study the effect of CⅡTA on the expression of HLA class Ⅱ molecules, the 5’-end cDNA of C Ⅱ TA gene was cloned from Raji cells by RT-PCR, Eukaryotic expression vector pcDNA-Ⅱ of sense RNA and HeLa cells induced by HLA Ⅱ class genes. The results showed that the expression of HLA-DR, DP and DQ in transfected pcDNA-Ⅱ cells was significantly lower than that in pcDNA3 empty vector induced by IFN-γ, while the expression of HLA class I molecules was not affected. Experiments show that C Ⅱ TA antisense RNA has a significant inhibitory effect on the expression of HLA class Ⅱ molecules, which provides a basis for further development of low immunogenicity of cell transplantation.